Collection of human genomic DNA from neonates: a comparison between umbilical cord blood and buccal swabs

Lehmann, Amalia S.; Haas, David M.; McCormick, Catherine L.; Skaar, Todd C.; Renbarger, Jamie L.

doi:10.1016/j.ajog.2010.12.013

Cited by 10 publications

(8 citation statements)

References 21 publications

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“…Genotyping was performed blind to phenotypic data. The average DNA yield was 36 μg ( SD 26 μg) which is comparable to yields reported in other studies using infant saliva (Lehmann, Haas, McCormick, Skaar, & Renbarger, ). Genotypes were recalled by two investigators and in the 2% where there were different calls due to weak bands on the gels, the samples were excluded.…”

Section: Methodssupporting

confidence: 83%

Evidence for interplay between genes and parenting on infant temperament in the first year of life: monoamine oxidase A polymorphism moderates effects of maternal sensitivity on infant anger proneness

Pickles

Hill

Breen

et al. 2013

Child Psychology Psychiatry

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Background:The low expression polymorphism of the MAOA gene in interaction with adverse environments (G 9 E) is associated with antisocial behaviour disorders. These have their origins in early life, but it is not known whether MAOA G 9 E occurs in infants. We therefore examined whether MAOA G 9 E predicts infant anger proneness, a temperamental dimension associated with later antisocial behaviour disorders. In contrast to previous studies, we examined MAOA G 9 E prospectively using an observational measure of a key aspect of the infant environment, maternal sensitivity, at a specified developmental time point. Methods: In a stratified epidemiological cohort recruited during pregnancy, we ascertained MAOA status (low vs. high expression alleles) from the saliva of 193 infants, and examined specific predictions to maternal report of infant temperament at 14 months from maternal sensitivity assessed at 29 weeks of age. Results: Analyses, weighted to provide general population estimates, indicated a robust interaction between MAOA status and maternal sensitivity in the prediction of infant anger proneness (p = .003) which became stronger once possible confounders for maternal sensitivity were included in the model (p = .0001). The interaction terms were similar in males (p = .010) and females (p = .016), but the effects were different as a consequence of an additional sex of infant by maternal sensitivity interaction. Conclusions: This prospective study provides the first evidence of moderation by the MAOA gene of effects of parenting on infant anger proneness, an important early risk for the development of disruptive and aggressive behaviour disorders. Keywords: Monoamine oxidase A, promoter polymorphism, maternal sensitivity, infant temperament, anger proneness, gene by environment interaction.

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Section: Methodssupporting

confidence: 83%

Evidence for interplay between genes and parenting on infant temperament in the first year of life: monoamine oxidase A polymorphism moderates effects of maternal sensitivity on infant anger proneness

Pickles

Hill

Breen

et al. 2013

Child Psychology Psychiatry

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“…The yields of gDNA extracted from whole blood and umbilical cords are comparable to previous studies where ∼6 μg gDNA/200 μl whole blood [28], ~100 μg gDNA/ 200 μl umbilical cord blood [29] were obtained (Table 2). The gDNA yield of DBS 180 ng/3.2 mm punch however was higher than previously published ~60 ng gDNA/3.2 mm punch [19] or 19-40 ng/3.0 mm punch [30].…”

Section: Discussionsupporting

confidence: 85%

Isolation of human genomic DNA for genetic analysis from premature neonates: a comparison between newborn dried blood spots, whole blood and umbilical cord tissue

et al. 2013

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BackgroundGenotyping requires biological sample collection that must be reliable, convenient and acceptable for patients and clinicians. Finding the most optimal procedure of sample collection for premature neonates who have a very limited blood volume is a particular challenge. The aim of the current study was to evaluate the use of umbilical cord (UC) tissue and newborn dried blood spot (DBS)-extracted genomic DNA (gDNA) as an alternative to venous blood-derived gDNA from premature neonates for molecular genetic analysis.All samples were obtained from premature newborn infants between 24-32 weeks of gestation. Paired blood and UC samples were collected from 31 study participants. gDNA was extracted from ethylenediaminetetraacetic acid (EDTA) anticoagulant-treated blood samples (~500 μl) and newborn DBSs (n = 723) using QIAamp DNA Micro kit (Qiagen Ltd., Crawley, UK); and from UC using Qiagen DNAeasy Blood and Tissue kit (Qiagen Ltd., Crawley, UK). gDNA was quantified and purity confirmed by measuring the A260:A280 ratio. PCR amplification and pyrosequencing was carried out to determine suitability of the gDNA for molecular genetic analysis. Minor allele frequency of two unrelated single nucleotide polymorphisms (SNPs) was calculated using the entire cohort.ResultsBoth whole blood samples and UC tissue provided good quality and yield of gDNA, which was considerably less from newborn DBS. The gDNA purity was also reduced after 3 years of storage of the newborn DBS. PCR amplification of three unrelated genes resulted in clear products in all whole blood and UC samples and 86%-100% of newborn DBS. Genotyping using pyrosequencing showed 100% concordance in the paired UC and whole blood samples. Minor allele frequencies of the two SNPs indicated that no maternal gDNA contamination occurred in the genotyping of the UC samples.ConclusionsgDNAs from all three sources are suitable for standard PCR and pyrosequencing assays. Given that UC provide good quality and quantity gDNA with 100% concordance in the genetic analysis with whole blood, it can replace blood sampling from premature infants. This is likely to reduce the stress and potential side effects associated with invasive sample collection and thus, greatly facilitate participant recruitment for genetic studies.

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“…(35) Although DNA quantity and quality are typically higher from cord blood compared with saliva or buccal cells, it is often difficult to obtain cord blood at the time of delivery from all subjects. (36) An increasing number of epigenetic studies have utilized saliva as a DNA source as it is readily available, can be followed serially, and is non-invasive. (35,37) DNA source was adjusted for in all of our multivariate models and results remained unchanged when DNA source was included in the models.…”

Section: Discussionmentioning

confidence: 99%

Epigenetic Variation in the Mu-Opioid Receptor Gene in Infants with Neonatal Abstinence Syndrome

Wachman

Hayes

Lester

et al. 2014

The Journal of Pediatrics

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Objective Neonatal abstinence syndrome (NAS) from in utero opioid exposure is highly variable with genetic factors appearing to play an important role. Epigenetic changes in cytosine:guanine (CpG) dinucleotide methylation can occur after drug exposure and may help to explain NAS variability. We correlated DNA methylation levels in the mu-opioid receptor (OPRM1) promoter in opioid-exposed infants and correlate them with NAS outcomes. Study design DNA samples from cord blood or saliva were analyzed for 86 infants being treated for NAS according to institutional protocol. Methylation levels at 16 OPRM1 CpG sites were determined and correlated with NAS outcome measures, including need for treatment, treatment with >2 medications, and length of hospital stay. We adjusted for co-variates and multiple genetic testing. Results Sixty-five percent of infants required treatment for NAS, and 24% required ≥2 medications. Hypermethylation of the OPRM1 promoter was measured at the −10 CpG in treated versus non-treated infants [adjusted difference δ=3.2% (95% CI 0.3–6.0%), p=0.03; NS after multiple testing correction]. There was hypermethylation at the −14 [δ=4.9% (95% CI 1.8–8.1%), p=0.003], −10 [δ=5.0% (95% CI 2.3–7.7%), p=0.0005)], and +84 [δ=3.5% (95% CI 0.6 – 6.4), p=0.02] CpG sites in infants requiring ≥2 medications which remained significant for −14 and −10 after multiple testing correction. Conclusions Increased methylation within the OPRM1 promoter is associated with worse NAS outcomes, consistent with gene silencing.

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Collection of human genomic DNA from neonates: a comparison between umbilical cord blood and buccal swabs

Cited by 10 publications

References 21 publications

Evidence for interplay between genes and parenting on infant temperament in the first year of life: monoamine oxidase A polymorphism moderates effects of maternal sensitivity on infant anger proneness

Evidence for interplay between genes and parenting on infant temperament in the first year of life: monoamine oxidase A polymorphism moderates effects of maternal sensitivity on infant anger proneness

Isolation of human genomic DNA for genetic analysis from premature neonates: a comparison between newborn dried blood spots, whole blood and umbilical cord tissue

Epigenetic Variation in the Mu-Opioid Receptor Gene in Infants with Neonatal Abstinence Syndrome

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